Programmable Polyproteams of Tyrosine Ammonia Lyases as Cross-Linked Enzymes for Synthesizing <i>p</i>-Coumaric Acid
Mingyu Jia,
Zhiyuan Luo,
Haomin Chen,
Bianqin Ma,
Li Qiao,
Qinjie Xiao,
Pengfei Zhang,
Anming Wang
Affiliations
Mingyu Jia
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Zhiyuan Luo
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Haomin Chen
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Bianqin Ma
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Li Qiao
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Qinjie Xiao
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Pengfei Zhang
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Anming Wang
Key Laboratory of Organosilicon Chemistry and Material Technology, College of Material, Chemistry and Chemical Engineering, Ministry of Education, Hangzhou Normal University, No. 2318, Road Yuhangtang, Hangzhou 311121, China
Ideal immobilization with enhanced biocatalyst activity and thermostability enables natural enzymes to serve as a powerful tool to yield synthetically useful chemicals in industry. Such an enzymatic method strategy becomes easier and more convenient with the use of genetic and protein engineering. Here, we developed a covalent programmable polyproteam of tyrosine ammonia lyases (TAL-CLEs) by fusing SpyTag and SpyCatcher peptides into the N-terminal and C-terminal of the TAL, respectively. The resulting circular enzymes were clear after the spontaneous isopeptide bonds formed between the SpyTag and SpyCatcher. Furthermore, the catalytic performance of the TAL-CLEs was measured via a synthesis sample of p-Coumaric acid. Our TAL-CLEs showed excellent catalytic efficiency, with 98.31 ± 1.14% yield of the target product—which is 4.15 ± 0.08 times higher than that of traditional glutaraldehyde-mediated enzyme aggregates. They also showed over four times as much enzyme-activity as wild-type TAL does and demonstrated good reusability, and so may become a good candidate for industrial enzymes.
